發(fā)布時(shí)間：2018-03-24 15:38

  本文選題：食管鱗狀細(xì)胞癌　切入點(diǎn)：MACC1　出處：《東南大學(xué)》2015年碩士論文

【摘要】：[研究目的]研究結(jié)腸癌轉(zhuǎn)移相關(guān)基因1 (Metastasis Associated In Colon Cancer 1, MACC1)對(duì)食管癌細(xì)胞增殖和遷移的作用及可能的機(jī)制。[研究方法]1.采用RT-PCR及Western blot檢測(cè)不同食管癌細(xì)胞株EC109、 EC9706、 TE13、 EC1和正常食管上皮細(xì)胞株HEEpiC中MACC1的表達(dá),選取低表達(dá)MACC1的細(xì)胞株TE13和高表達(dá)MACC1的細(xì)胞株EC109進(jìn)行后續(xù)實(shí)驗(yàn)。2.構(gòu)建攜帶MACC1的腺病毒載體Ad-MACC1和MACC1 siRNA, Ad-MACC1感染TE13細(xì)胞,增加MACC1的表達(dá),MACC1 siRNA轉(zhuǎn)染EC109食管癌細(xì)胞,降低MACC1的表達(dá),通過Western blot檢測(cè)感染或轉(zhuǎn)染效果。3. Ad-MACC1和MACC1 siRNA分別處理TE13和EC109細(xì)胞后,通過MTT實(shí)驗(yàn)檢測(cè)MACC1對(duì)人食管癌細(xì)胞增殖的影響,流式細(xì)胞術(shù)檢測(cè)對(duì)MACC1細(xì)胞周期的影響。采用劃痕實(shí)驗(yàn)及克隆形成實(shí)驗(yàn)觀察感染或轉(zhuǎn)染前后食管癌細(xì)胞侵襲及遷移變化。4.采用Western blot檢測(cè)Ad-MACC1和MACC1 siRNA處理食管癌細(xì)胞TE13和EC109后的增殖細(xì)胞核抗原(PCNA)和c-MET等的表達(dá)。[研究結(jié)果]1. MACC1在食管癌細(xì)胞株EC9706和Eca109中表達(dá)較正常食管上皮細(xì)胞株HEEpiC中高,而在TE13和EC1中表達(dá)較低。2.成功構(gòu)建Ad-MACC1和MACC1 siRNA, Ad-MACC1感染TE13細(xì)胞后,MACC1基因和蛋白的表達(dá)增加。而MACC1 siRNA轉(zhuǎn)染EC109細(xì)胞后,MACC1基因和蛋白的表達(dá)降低。3. Ad-MACC1感染TE13細(xì)胞后,細(xì)胞的增殖和遷移明顯的增加。而MACC1 siRNA轉(zhuǎn)染EC109細(xì)胞后,細(xì)胞的增殖和遷移受到明顯的抑制。4. Ad-MACC1感染TE13細(xì)胞后,PCNA和c-MET基因和蛋白表達(dá)明顯增加,而MACC1 siRNA轉(zhuǎn)染EC109細(xì)胞后,PCNA和c-MET基因和蛋白表達(dá)較陰性對(duì)照組降低。[結(jié)論]MACC1在EC9706和Eca109中表達(dá)較TE13和EC1高,其高表達(dá)可促進(jìn)細(xì)胞的增殖和遷移,而表達(dá)下調(diào)后明顯抑制食管癌細(xì)胞的增殖、侵襲及轉(zhuǎn)移潛能,MACC1高表達(dá)促進(jìn)PCNA和c-MET的表達(dá),而低表達(dá)抑制其表達(dá)。MACC1促進(jìn)食管癌細(xì)胞的增殖、侵襲及轉(zhuǎn)移。其機(jī)制可能通過調(diào)節(jié)下游基因c-MET表達(dá)水平實(shí)現(xiàn)。
[Abstract]:[objective] to study the effect of Metastasis Associated in Colon Cancer 1 (MACC1) on the proliferation and migration of esophageal carcinoma cells and its possible mechanism. 1. To detect different esophageal cancer cell lines EC109, EC9706, TE13 by RT-PCR and Western blot. Expression of MACC1 in EC1 and normal esophageal epithelial cell line HEEpiC. The cell line TE13 with low expression of MACC1 and the cell line EC109 with high expression of MACC1 were selected for further experiment. 2. The adenovirus vectors Ad-MACC1 and MACC1 siRNAs carrying MACC1 were constructed, Ad-MACC1 infected TE13 cells, and the expression of MACC1 was increased. MACC1 siRNA was transfected into EC109 esophageal carcinoma cells, and the expression of MACC1 was decreased. Western blot was used to detect the effect of infection or transfection. Ad-MACC1 and MACC1 siRNA were used to treat TE13 and EC109 cells respectively. The effect of MACC1 on the proliferation of human esophageal carcinoma cells was detected by MTT assay. The effect of flow cytometry on the cell cycle of MACC1. The invasion and migration of esophageal carcinoma cells before and after infection or transfection were observed by scratch test and clone formation assay. 4. The TE13 of esophageal cancer cells treated with Ad-MACC1 and MACC1 siRNA were detected by Western blot. Expression of proliferating cell nuclear antigen (PCNA) and c-MET after EC109. [results] 1.The expression of MACC1 in EC9706 and Eca109 cells was higher than that in normal esophageal epithelial cell line HEEpiC. Ad-MACC1 and MACC1 siRNAs were successfully constructed in TE13 and EC1. The expression of MACC1 gene and protein increased after Ad-MACC1 infection in TE13 cells, while the expression of MACC1 gene and protein in EC109 cells transfected with MACC1 siRNA decreased .3. Ad-MACC1 infected TE13 cells. The proliferation and migration of EC109 cells were significantly increased after transfection of MACC1 siRNA, and the proliferation and migration of EC109 cells were inhibited significantly. 4. The expression of c-MET gene and protein in TE13 cells infected with Ad-MACC1 was significantly increased. The expression of MACC1 and c-MET gene and protein in EC109 cells transfected with MACC1 siRNA were lower than those in negative control group. [conclusion] the expression of MACC1 in EC9706 and Eca109 was higher than that in TE13 and EC1, and the high expression of MACC1 could promote the proliferation and migration of EC109 cells. The down-regulated expression of MACC1 significantly inhibited the proliferation of esophageal cancer cells, and the high expression of MACC1 promoted the expression of PCNA and c-MET, while the low expression of MACC1 inhibited the proliferation of esophageal cancer cells. Invasion and metastasis. The mechanism may be through regulation of downstream gene c-MET expression level.
【學(xué)位授予單位】：東南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R735.1

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